Procedures for Determination of Detection Limits Application to High-performance Liquid Chromatography Analysis of Fat-soluble Vitamins in Human Serum
Procedures for Determination of Detection Limits Application to High-performance Liquid Chromatography Analysis of Fat-soluble Vitamins in Human Serum
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DOI：
10.1097/ede.0b013e3181ce9a61
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发表时间：
2010-07-01
期刊：
影响因子：
5.4
通讯作者：
Whitcomb, Brian W.
中科院分区：
文献类型：
--
作者：
Browne, Richard W.;Whitcomb, Brian W.
背景资料：实验室数据分析中的问题通常出现在流行病学研究中，其中使用的生物标志物的检测阈值较低。存在各种阈值，包括检测限（LOD）、定量限（LOQ）和空白限（LOB）。选择适当的策略来处理受这些限制影响的数据依赖于对检测限及其确定的性质的正确理解。在本文中，我们展示了实验和统计程序一般用于估计不同的检测限，根据标准程序的背景下，分析脂溶性维生素和微量营养素在人体serum.Methods：脂溶性维生素和微量营养素进行了分析，高效液相色谱二极管阵列检测。重复分析模拟血清基质空白，通过使用观察到的空白分布进行参数化分析，以及通过使用秩进行非参数化分析，以确定LOB。通过将LOB相关信息与稀释至低水平（从LOB至2-3倍LOB）的标准参比物质（SRM）的重复分析数据相结合，确定LOD。通过绘制SRM重复样品的相对标准偏差（RSD）与浓度的关系曲线，实验确定LOQ，其中LOQ为RSD为20%时的浓度。结果：给出了确定LOB、LOD和LOQ的实验方法和示例统计程序。这些数量报告为每个测量的分析物。结论：对于许多分析，有相当多的信息低于LOQ。流行病学研究必须了解这些检测限的性质，以及如何估计它们以适当处理受影响的数据。
Background: Problems in the analysis of laboratory data commonly arise in epidemiologic studies in which biomarkers subject to lower detection thresholds are used. Various thresholds exist including limit of detection (LOD), limit of quantification (LOQ), and limit of blank (LOB). Choosing appropriate strategies for dealing with data affected by such limits relies on proper understanding of the nature of the detection limit and its determination. In this paper, we demonstrate experimental and statistical procedures generally used for estimating different detection limits according to standard procedures in the context of analysis of fat-soluble vitamins and micronutrients in human serum.Methods: Fat-soluble vitamins and micronutrients were analyzed by high-performance liquid chromatography with diode array detection. A simulated serum matrix blank was repeatedly analyzed for determination of LOB parametrically by using the observed blank distribution as well as nonparametrically by using ranks. The LOD was determined by combining information regarding the LOB with data from repeated analysis of standard reference materials (SRMs), diluted to low levels; from LOB to 2-3 times LOB. The LOQ was determined experimentally by plotting the observed relative standard deviation (RSD) of SRM replicates compared with the concentration, where the LOQ is the concentration at an RSD of 20%.Results: Experimental approaches and example statistical procedures are given for determination of LOB, LOD, and LOQ. These quantities are reported for each measured analyte.Conclusion: For many analyses, there is considerable information available below the LOQ. Epidemiologic studies must understand the nature of these detection limits and how they have been estimated for appropriate treatment of affected data.